Compositions and methods for stabilizing biological molecules upon lyophilization

Abstract

The present invention is directed to compositions and methods for the preparation of biological molecules, in particular MenC-CRM 197. Biological molecules dissolved in dissolution buffers comprising at least one amorphous excipient and an amorphous buffer exhibit reduced aggregation and proteolytic breakdown.

We claim:  
     
         1 . A dissolution buffer comprising at least one amorphous excipient, at least one amorphous buffer, and at least one MenC immunogen.  
     
     
         2 . The dissolution buffer of  claim 1  wherein the amorphous buffer is an organic buffer.  
     
     
         3 . The dissolution buffer of  claim 2  wherein the organic buffer is selected from the group consisting of imidazole buffer and histidine buffer.  
     
     
         4 . The dissolution buffer of  claim 1  wherein the amorphous excipient is a sugar.  
     
     
         5 . The dissolution buffer of  claim 4  wherein the sugar is sucrose.  
     
     
         6 . The dissolution buffer of  claim 1  wherein the MenC immunogen is MenC-CRM 197.  
     
     
         7 . A method for stabilizing one or more MenC immunogens upon lyophilization comprising; 
 (a) dissolving the MenC immunogen in a dissolution buffer comprising at least one amorphous excipient and an amorphous buffer to form a mixture, and    (b) lyophilizing the mixture.    
     
     
         8 . The method of  claim 7  wherein the MenC immunogen is MenC-CRM 197.  
     
     
         9 . A lyophilized composition stabilized according to the method of  claim 7  comprising at least one amorphous excipient, at least one amorphous buffer, and at least one MenC immunogen.  
     
     
         10 . A method for storing one or more MenC immunogens comprising; 
 (a) dissolving the MenC immunogen in a dissolution buffer comprising at least one amorphous excipient and an amorphous buffer to form a mixture, and    (b) lyophilizing the mixture.    
     
     
         11 . The method of  claim 10  wherein the MenC immunogen is MenC-CRM 197.  
     
     
         12 . A method for lyophilization of a MenC immunogen comprising; 
 dissolving a MenC immunogen in a dissolution buffer comprising at least one amorphous excipient and an amorphous buffer to form a mixture;    loading the mixture from about 10° C. to about 20° C. under from about 1000 Pa to about 2000 Pa of pressure;    freezing the mixture from about 1 hour to about 4 hours to a temperature from about −70° C. to about −50° C. under from about 1000 Pa to about 2000 Pa of pressure;    drying the mixture for from about 18 hours to about 30 hours at a temperature from about −50° C. to about −35° C. under a pressure from about 5 Pa to about 10 Pa; and    drying the mixture for from 2 hours to about 12 hours at a temperature from about 10° C. to about 20° C. under a pressure from about 5 Pa to about 10 Pa.    
     
     
         13 . The method of  claim 12  wherein the MenC immunogen is MenC-CRM 197.  
     
     
         14 . A method for lyophilization of a MenC immunogen comprising; 
 dissolving a MenC immunogen in a dissolution buffer comprising at least one amorphous excipient and an amorphous buffer to form a mixture;    loading the mixture at about 10° C. under 1000 Pa of pressure;    freezing the mixture for about 2 hours to a temperature of about −50° C. under about 1000 Pa of pressure;    drying the mixture for about 24 hours at a temperature of about −35° C. under a pressure of about 5 Pa; and    drying the mixture for about 8 hours at a temperature of about 20° C. under a pressure of about 5 Pa.    
     
     
         15 . The method of  claim 14  wherein the immunogen is Men C-CRM 197.  
     
     
         16 . A method for stabilizing a MenC vaccine upon lyophilization comprising; 
 (a) dissolving the MenC vaccine in a dissolution buffer comprising at least one amorphous excipient and an amorphous buffer to form a mixture, and    (b) lyophilizing the mixture.    
     
     
         17 . The method of  claim 16  wherein the MenC vaccine is MenC-CRM 197.  
     
     
         18 . A MenC vaccine stabilized according to the method of  claim 16  comprising at least one amorphous excipient and at least one amorphous buffer.